Curators

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Abstract

Lyse red blood cells while leaving the Plasmodium falciparum parasite intact with it's parasite membrane and parasitophorous vacoule membrane. Typically used right before freezing down parasites for genomic DNA extraction, or for getting rid of hemoglobin right before running a Western Blot on parasite extracts.

Reagents

0.15% Saponin in PBS

1X PBS

12 mL of Plasmodium falciparum blood culture at 4% hematocrit

Equipment

Refrigerated microcentrifuge

Procedure

Centrifuge parasite culture at 1400 rpm (~484xg) for 3 minutes.

Aspirate media.

Resuspend pellet in 1 ml of 0.15% Saponin (in aliquots in freezer) and transfer to a 1.5 ml eppendorf tube.

Incubate for 5 minutes on ice and vortex each minute.

Spin at 6000 rpm for 3 minutes at 4 degrees C.

Wash with 1 ml 1X PBS (spin at 6000 rpm for 3 minutes) 3 to 4 times.

Each wash step consists of aspirating the supernatant and resuspending the pellet with new wash buffer, before centrifuging.

Aspirate off last wash.

Store at -20 degrees C until ready to use.

Critical steps

Referenced from the main protocol, a more thorough explanation of particularly important steps in the protocol.

Troubleshooting

Referenced from the main protocol, an explanation of what can cause things to go wrong with the protocol.

Notes

Referenced from the main protocol, any comments about the protocol should be made here; i.e. how it was developed. Any comments added should be signed (by adding *'''~~~~''': in front) and explained. Links to FAQs/tips provided by other sources such as the manufacturer or other websites would be best made here.
Anecdotal observations that might be of use to others can also be posted here; e.g. 'my cells were still floating'.

It might also be good to add an image to show the workflow and timescales for experiment planning.

Acknowledgments

Acnkowledge any help you had in development, testing, writing this protocol.